KR102012416B1 - Antimicrobial composition including extract of schizophyllum commune strain culture and preparing method thereof - Google Patents

Abstract

The present application relates to an antimicrobial composition containing the skirt mushroom strain culture extract and a method for preparing the same.

Description

ANTIMICROBIAL COMPOSITION INCLUDING EXTRACT OF SCHIZOPHYLLUM COMMUNE STRAIN CULTURE AND PREPARING METHOD THEREOF}

The present application relates to an antimicrobial composition containing the skirt mushroom strain culture extract and a method for preparing the same.

As the term “well being” or LOHAS (lifestyle of health and sustainability) is widely used in recent years, there is a growing movement around the world to simultaneously pursue and practice healthy living and environmental preservation. Among them, dietary life, especially agricultural products, are sensitive to these changes because they are strongly related to health and take effect directly by ingesting them directly. Korea also has been concerned about environmental disasters such as yellow dust, mad cow disease, foot-and-mouth disease, and environmental pollution since the 2000s. Consumers' interest in safe food is growing. As a result, preference for eco-friendly agricultural products increases, and interest and research on biological control such as biological pesticides for eco-friendly agricultural products are increasing.

Existing methods of controlling pathogenic fungi in crops include partial resistance varieties, cultivation and the use of chemical antifungal agents. However, new varieties may take a long time, and the pathogens may adapt and change the resistance of plants, resulting in a short-lived beneficial effect, and rotation or soil solar therapy is difficult to implement in large-scale agriculture. In addition, chemical antifungal agents are reported to be less effective for the cost and to adversely affect the environment and human health. Among them, chemical antifungal agents (pesticides) have been important in agricultural production in terms of improving the yield and quality of crops and increasing supply according to population growth. Until now, chemical control through chemical fertilizers and organic synthetic pesticides Done. However, the adverse effects of hardly decomposable residual toxicity due to overuse of pesticides pollute air, soil, and groundwater, and adverse effects on the environment and ecosystem, and decrease in medicinal effects due to acquiring pesticide resistance of phytopathogens. As a biological control method using microbial agents against these side effects, as an effective and sustainable alternative to plant diseases, Bacillus spp., Pseudomonas spp., Streptomyces spp. And non-pathogenic Pusa Research on the control of soil-derived plant pathogens with microorganisms such as Fusarium spp. Has been conducted. Biological control methods have been highlighted as effective countermeasures because they have no residual toxicity and do not disturb agricultural ecosystems. However, the share of biological control is low in the global pesticide market.

Mushrooms, low-calorie health foods rich in dietary fiber and protein, are used primarily for food and medicinal purposes. Mushrooms have been reported to produce various functional substances such as antibiotics, antiviral substances, as well as biological functions such as anti-cancer, anti-cholesterol substances, blood pressure regulators, immunomodulators. Therefore, mushrooms are considered to be highly applicable as a substitute for chemical pesticides.

On the other hand, the skirt mushroom belongs to the skirt mushroom family, and the shade is fan-shaped or skirt-shaped about 1 cm to 3 cm in diameter, and the surface is densely formed with coarse hair of white, gray or grey-brown. The girth is divided by the number of wrinkles. The fruiting bodies are leathery and contract when dry or wet. Wrinkles are white or grayish white. Wrinkle ends are soft and double. There is no stand and a part of the lamp lives on the host. Spores are 5-6 μm × 4-5 μm subspheres with smooth and light surface. Spores are flesh-colored. The cysts are fusiform, without horns on the top, 60-80 μm in length. It is a fungi mushroom that grows or overlaps with dead trees or living bark all four seasons. Occurs from spring to autumn, forming clusters in overlapping groups. Skirt mushrooms are keratinous mushrooms, which are difficult to use for food. However, various examples have been reported for extracting polysaccharides from liquid culture mycelium or culture medium of purely isolated mycelium.

Korean Patent Laid-Open No. 10-2012-0091680 discloses Streptomyces sp. A1009 and its culture method having excellent antibacterial activity against phytopathogens.

The present invention is to solve the above-mentioned problems of the prior art, an object of the present invention is to provide an antimicrobial composition and method for producing the same effective for the control and prevention of plant diseases.

However, the technical problem to be achieved by the embodiments of the present application is not limited to the technical problems as described above, and other technical problems may exist.

As a technical means for achieving the above technical problem, the first aspect of the present application provides an antimicrobial composition comprising a chia mushroom strain culture extract containing schizostatin or derivatives thereof.

According to one embodiment of the present application, the schizstatin or derivatives thereof may include a material represented by the following Chemical Formula 1, but is not limited thereto;

[Formula 1]

[Wherein, R ₁ to R ₃ are each independently hydrogen, oxygen, hydroxyl group, substituted or unsubstituted alkyl group, substituted or unsubstituted alkenyl group, substituted or unsubstituted alkynyl group, substituted or unsubstituted cycloalkyl group, substituted Or an unsubstituted alkoxy group, a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted alkoxycarbonyl group, and n is a number from 1 to 6.

According to one embodiment of the present application, the schizstatin or derivatives thereof may include a substance represented by the following Chemical Formula 2, but is not limited thereto;

[Formula 2]

.

.

According to one embodiment of the present application, the schizstatin or a derivative thereof may include a material represented by the following Chemical Formula 3, but is not limited thereto;

[Formula 3]

.

.

According to one embodiment of the present application, the chizostatin or a derivative thereof may include a material represented by the following Chemical Formula 4, but is not limited thereto;

[Formula 4]

.

.

According to one embodiment of the present application, the schizstatin or a derivative thereof may include a material represented by the following Chemical Formula 5, but is not limited thereto;

[Formula 5]

.

.

According to the exemplary embodiment of the present application, the schizostatin or a derivative thereof may include a material represented by the following Chemical Formula 6, but is not limited thereto;

[Formula 6]

.

.

According to a second aspect of the present disclosure, a step of culturing a skirt mushroom strain on a medium; Obtaining a first extract from the cultured skirt mushroom strain using a first solvent; And extracting chizostatin or a derivative thereof from the first extract using a second solvent.

According to one embodiment of the present application, the medium is PDA (potato dextrose agar), PDB (potato dextrose broth), SDA (sabouraud dextrose agar), DSA (dextrose starch agar), MM (minimal medium), CM (complete medium) , CDA (czapek dox agar) or CMC (carboxyl methyl cellulose) may be selected from, but is not limited thereto.

According to one embodiment of the present application, the first solvent may be selected from the group consisting of water, acetone, C ₁ -C ₄ alcohol or a mixed solvent thereof, and combinations thereof, but is not limited thereto.

According to one embodiment of the present application, the second solvent may include an organic solvent, but is not limited thereto.

According to one embodiment of the present application, the extraction of the chyzostatin or derivatives thereof may be performed by extraction, distribution of water, or chromatography, but is not limited thereto.

The above-mentioned means for solving the problems are merely exemplary and should not be construed as limiting the present application. In addition to the above-described exemplary embodiments, additional embodiments may exist in the drawings and detailed description of the invention.

According to the aforementioned problem solving means of the present application, the present application can provide an antimicrobial composition and a method for producing the same.

The antimicrobial composition according to the present invention can control and prevent plant diseases by showing an antifungal effect on phytopathogenic fungi as a compound derived from natural materials.

In particular, it was confirmed the antifungal effect of the novel compounds isolated from the mushroom mushroom culture medium, it is possible to use it for the control and prevention of fungal diseases.

1 is a flow chart of a method for producing an antimicrobial composition according to one embodiment of the present application.
Figure 2 is a microscope image showing the antimicrobial activity of the antimicrobial composition according to an embodiment of the present application.

DETAILED DESCRIPTION Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present disclosure.

As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention. In the drawings, parts irrelevant to the description are omitted for simplicity of explanation, and like reference numerals designate like parts throughout the specification.

Throughout this specification, when a portion is "connected" to another portion, this includes not only "directly connected" but also "electrically connected" with another element in between. do.

Throughout this specification, when a member is said to be located on another member "on", "upper", "top", "bottom", "bottom", "bottom", this means that any member This includes not only the contact but also the presence of another member between the two members.

Throughout this specification, when a part is said to "include" a certain component, it means that it can further include other components, without excluding the other components unless specifically stated otherwise.

As used herein, the terms "about", "substantially", and the like, are used at, or in close proximity to, numerical values when manufacturing and material tolerances inherent in the meanings indicated are provided to aid the understanding herein. In order to prevent the unfair use of unscrupulous infringers. In addition, throughout this specification, "step to" or "step of" does not mean "step for."

Throughout this specification, the term "combination of these" included in the expression of the makushi form means one or more mixtures or combinations selected from the group consisting of constituents described in the expression of the makushi form, wherein the constituents It means to include one or more selected from the group consisting of.

Throughout this specification, description of "A and / or B" means "A, B, or A and B."

Examples of the substituents are described below, but are not limited thereto.

As used herein, the term "substituted or unsubstituted" is hydrogen; Hydroxyl group; Alkyl groups; Alkenyl groups; Alkynyl group; Cycloalkyl group; An alkoxy group; Heterocyclic group; Aralkyl group; Aryloxy group; An alkoxycarbonyl group; means substituted or unsubstituted with one or more substituents selected from the group consisting of, or substituted or unsubstituted two or more substituents of the substituents exemplified above. For example, "a substituent to which two or more substituents are linked" may be a heterocyclic group substituted with an alkyl group or an alkyl group substituted with a heterocyclic group.

In the present specification, an alkyl group means a saturated aliphatic hydrocarbon group containing 1 to 20 (eg, 2 to 18, 3 to 18, 1 to 8, 1 to 6, 1 to 4, or 1 to 3) carbon atoms. do. The alkyl group can be straight chain, branched chain, cyclic or any combination thereof. Specific examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl or 2-ethylhexyl. Alkyl groups may be substituted (ie, optionally substituted) with one or more substituents or may be multicyclic as described below.

In the present specification, an alkenyl group means an aliphatic carbon group containing 2 to 20 (eg, 2 to 18, 2 to 8, 2 to 6, or 2 to 4) carbon atoms and at least one double bond. do. Like alkyl groups, alkenyl groups can be straight, branched or cyclic or any combination thereof. Specific examples of alkenyl groups include, but are not limited to, allyl, isoprenyl, 2-butenyl, and 2-hexenyl. Alkenyl groups may be optionally substituted with one or more substituents as described below.

As used herein, an alkynyl group refers to an aliphatic carbon group containing 2 to 20 (eg, 2 to 8, 2 to 6 or 2 to 4) carbon atoms and having at least one triple bond. Alkynyl groups may be straight chain, branched or cyclic or any combination thereof. Specific examples of alkynyl groups include, but are not limited to, propargyl and butynyl. Alkynyl groups may be optionally substituted with one or more substituents as described below.

In the present specification, the cycloalkyl group is not particularly limited, but is preferably 3 to 60 carbon atoms (for example, 3 to 30, 3 to 20, 3 to 6). Specific examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, 3-methylcyclopentyl, 2,3-dimethylcyclopentyl, cyclohexyl, 3-methylcyclohexyl, 4-methylcyclohexyl, 2,3-dimethyl Cyclohexyl, 3,4,5-trimethylcyclohexyl, 4-tert-butylcyclohexyl, cycloheptyl, cyclooctyl.

In the present specification, the heterocyclic group is a heterocyclic group including one or more of N, P, O, S, Se, Ge, and Si as heteroatoms, and the carbon number is not particularly limited, but is preferably 2 to 60 carbon atoms. Specific examples of the heterocyclic group include thiophene group, furan group, pyrrole group, imidazole group, thiazole group, oxazole group, oxadiazole group, triazole group, pyridyl group, bipyridyl group, pyrimidine group, triazine group, triazole group , Acridine group, carboline group, acenaphthopyrazine group, acenaphthoquinoxaline group, indenoquinazolin, indenoisoquinoline group, indenoquinoline group, pyridoindole group, pyridazine group, pyrazine group, quinoline Nyl group, quinazoline group, quinoxaline group, phthalazine group, pteridine group, pyrido pyrimidine group, pyrido pyrazine group, pyrazino pyrazine group, isoquinoline group, indole group, carbazole group, benzoxazole group, benzoimimi Diazole group, benzothiazole group, benzocarbazole group, dibenzocarbazole group, benzothiophene group, dibenzothiophene group, benzofuran group, dibenzofuran group, phenanthroline group, thiazolyl group, isoxazolyl Group, oxadiazolyl group, thiadiazolyl group, benzothiazole Groups, phenoxazinyl groups, phenothiazinyl groups, dibenzofuranyl groups, benzoquinazolins, pyridopyrimidine groups, indazole groups, benzoferimidinone groups, benzoferimidine groups, pyridoindole groups, and the like, but are limited thereto. It is not. The heterocyclic group includes an aliphatic heterocyclic group and an aromatic heterocyclic group.

As used herein, an aryl group used alone or as part of a large moiety, such as in an aralkyl group, an alkoxy group or an aryloxy group, is a monocyclic (eg phenyl), bicyclic (eg indenyl, Naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl), and tricyclic (eg, fluorenyl tetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systems, wherein the monocyclic The system is aromatic or at least one of the rings in a bicyclic or tricyclic system is aromatic. Bicyclic and tricyclic groups include benzofused 2-3 membered carbocyclic rings. For example, benzofused groups include phenyl fused with two or more C _4-8 carbocyclic moieties. Aryl may be optionally substituted with one or more substituents as described below.

As used herein, an alkoxycarbonyl group refers to -COOR, where R is alkyl as defined above, for example, methoxycarbonyl, ethoxycarbonyl and the like.

In the present specification, an alkoxy group refers to an alkyl-O— group, where “alkyl” is as previously defined.

In the present specification, "antibacterial" or "antimicrobial activity" means a property that resists microorganisms, such as bacteria and fungi, more specifically, antibiotics and the like means properties that inhibit the growth or proliferation of pathogenic fungi. do.

In the present specification, the "antimicrobial composition" is a composition having an activity that inhibits the growth of microorganisms such as bacteria or fungi, and may include all forms used in various fields requiring antimicrobial effects, for example, drugs , Quasi-drugs, food additives or feed additives. Specifically, for medicines, such as antibiotics and anti-fouling agents, for food preservation and antibacterial purposes, for agriculture, for the purpose of antibacterial, sterilization, disinfection, cosmetics and household goods for dandruff control, athlete's foot prevention, armpit collection It may be used in products directly related to microorganisms, such as inhibitory and anti-acne, or may be used for antiseptic, antibacterial or sterilizing purposes such as cleaning detergents or dish washing detergents, but is not limited thereto.

Hereinafter, the antimicrobial composition of the present application and a method for preparing the same will be described in detail with reference to embodiments, examples, and drawings. However, the present application is not limited to these embodiments, examples and drawings.

A first aspect of the present disclosure relates to an antimicrobial composition comprising a skirt mushroom strain culture extract containing schizostatin or a derivative thereof.

The skirt mushroom belongs to the skirt mushroom family, and the shade is fan-shaped or skirt-shaped about 1 cm to 3 cm in diameter, and its surface is densely formed with coarse hair of white, gray or grey-brown. The girth is divided by the number of wrinkles. The fruiting bodies are leathery and contract when dry or wet. Wrinkles are white or grayish white. Wrinkle ends are soft and double. There is no stand and a part of the lamp lives on the host. Spores are 5-6 µm × 4-5 µm subspheres with smooth and light surface. Spores are flesh-colored. The cysts are fusiform, without horns on the top, 60-80 μm in length. It is a fungi mushroom that grows or overlaps with dead trees or living bark all four seasons. Occurs from spring to autumn, forming clusters in overlapping groups. Compounds showing various activities have been found from the Schizophytes, for example, EPS (exopolysaccharide) derived from the anti-inflammatory activity, schizostatin, a squalene synthase inhibitor, and schizophyllan with anticancer activity ( schizophyllan) has been found.

According to one embodiment of the present application, the schizstatin or derivatives thereof may include a material represented by the following Chemical Formula 1, but is not limited thereto;

[Formula 1]

.

.

In formula, R <_1> -R <_3> is respectively independently hydrogen, oxygen, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, a substituted or unsubstituted alkynyl group, a substituted or unsubstituted cycloalkyl group, substituted or An unsubstituted alkoxy group, a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted alkoxycarbonyl group, and n may be 1 to 6, but It is not limited.

According to one embodiment of the present application, the schizstatin or derivatives thereof may include a substance represented by the following Chemical Formula 2, but is not limited thereto;

[Formula 2]

.

.

The material represented by Chemical Formula 2 is white powder, has a molecular formula of C ₂₀ H ₃₀ O ₄ , and has a molecular weight of 334.

The material represented by Chemical Formula 2 has a mass spectrometry spectrum of EI-MS m / z 333.0 [MH] ⁻ .

The material represented by Chemical Formula 2 is ppm (600 MHz, CD ₃ OD) 6.72 (s, H-2), 2.79 (t, J = 7.56, H-3), 2.18 (m, H-5), 5.16 ( t, J = 7.56, H-6), 1.59 (s, 7-CH ₃ ), 1.97 (t, H-8), 2.07 (m, H-9), 5.11 (t, J = 6.87, H-10 ), 1.59 (s, 11-CH ₃ ), 1.97 (t, H-12), 2.07 (m, H-13), 5.09 (t, J = 6.87, H-14), 1.59 (s, 15-CH ₃ ), the hydrogen nuclear magnetic resonance spectrum of 1.66 (s, H-16) is shown.

The substance represented by Chemical Formula 2 is ppm (600 MHz, CD ₃ OD) 169.1 (C-1), 128.4 (C-2), 148.7 (C-3), 170.2 (3-COOH), 28.9 (C-4 ), 28.8 (C-5), 124.5 (C-6), 137.4 (C-7), 16.2 (7-CH ₃ ), 40.9 (C-8), 27.8 (C-9), 125.5 (C-10 ), 136.0 (C-11), 16.3 (11-CH ₃ ), 41.0 (C-12), 27.9 (C-13), 125.6 (C-14), 132.2 (C-15), 17.9 (15-CH ₃ ) and a carbon nuclear magnetic resonance spectrum of 26.1 (C-16).

The substance represented by the above formula (2) was identified as schizostatin by the above results.

According to one embodiment of the present application, the schizstatin or a derivative thereof may include a material represented by the following Chemical Formula 3, but is not limited thereto;

[Formula 3]

.

.

The substance represented by Chemical Formula 3 is a colorless oil, has a molecular formula of C ₂₁ H ₃₆ O ₆ , and has a molecular weight of 384.

The material represented by Chemical Formula 3 has a mass spectrometry spectrum of EI-MS m / z 407.0 [M + Na] ⁺ .

The material represented by Chemical Formula 3 is ppm (600 MHz, CD ₃ OD) 5.23, 5.16 (dd, J = 11, 17.2, H-1), 6.03 (dd, J = 11, 17.9, H-2), 1.33 (s, 3-CH ₃ ), 1.64 (t, J = 8.25, H-4), 2.07 (m, H-5), 5.11 (t, J = 6.87, H-6), 1.60 (s, 7- CH ₃ ), 1.97 (t, J = 8.25, H-8), 2.04 (m, H-9), 5.09 (t, J = 6.87, H-10), 1.60 (s, 11-CH ₃ ), 1.67 (s, H-12), 4.59 (s, H-1 '), 3.75 (d, J = 3.4, H-2'), 3.43 (dd, J = 3.4, 9.28, H-3 '), 3.57 ( t, J = 9.28, H-4 '), 3.11 (m, H-5'), 3.69, 3.80 (m, H-6 ').

The substance represented by Chemical Formula 3 is ppm (600 MHz, CD ₃ OD) 115.1 (C-1), 144.4 (C-2), 81.3 (C-3), 23.9 (3-CH ₃ ), 41.4 (C- 4), 27.9 (C-5), 125.9 (C-6), 136.1 (C-7), 16.3 (7-CH ₃ ), 40.9 (C-8), 23.7 (C-9), 125.6 (C- 10), 132.2 (C-11), 17.9 (11-CH ₃ ), 26.0 (C-12), 96.6 (C-1 '), 74.2 (C-2'), 75.7 (C-3 '), 66.5 Carbon nuclear magnetic resonance spectra of (C-4 '), 78.0 (C-5') and 63.0 (C-6 ') are shown.

The substance represented by the said Formula (3) was identified as the novel compound SC-3 by the said result.

According to one embodiment of the present application, the chizostatin or a derivative thereof may include a material represented by the following Chemical Formula 4, but is not limited thereto;

[Formula 4]

.

.

The material represented by Chemical Formula 4 is colorless oil, has a molecular formula of C ₁₅ H ₂₆ O, and has a molecular weight of 222.

The material represented by Chemical Formula 4 has a mass spectrometry spectrum of EI-MS m / z 245.1 [M + Na] ⁺ .

The material represented by Chemical Formula 4 is ppm (600 MHz, CD ₃ OD) 5.19 (dd, J = 1.37, 17.53, H-1), 5.91 (dd, J = 11, 17.18, H-2), 1.25 (s , 3-CH ₃ ), 1.50 (m, H-4), 2.06 (m, H-5), 5.10 (m, H-6), 1.60 (s, 7-CH ₃ ), 1.97 (m, H- 8), hydrogen nuclear magnetic resonance of 2.02 (m, H-9), 5.03 (dd, J = 1.37, 11, H-10), 1.60 (s, 11-CH ₃ ), 1.67 (s, H-12) It shows the spectrum.

The material represented by Chemical Formula 4 is ppm (600 MHz, CD ₃ OD) 112.1 (C-1), 146.5 (C-2), 74.0 (C-3), 27.7 (3-CH ₃ ), 43.6 (C- 4), 27.9 (C-5), 125.9 (C-6), 136.0 (C-7), 16.1 (7-CH ₃ ), 41.0 (C-8), 23.8 (C-9), 125.6 (C- 10), 132.3 (C-11), 17.9 (11-CH ₃ ), and 26.0 (C-12).

The material represented by Chemical Formula 4 was identified as a novel compound nerodilol by the above results.

According to one embodiment of the present application, the schizstatin or a derivative thereof may include a material represented by the following Chemical Formula 5, but is not limited thereto;

[Formula 5]

.

.

The substance represented by Chemical Formula 5 is colorless oil, has a molecular formula of C ₁₇ H ₂₆ O ₅ , and has a molecular weight of 310.

The material represented by Chemical Formula 5 has a mass spectrometry spectrum of EI-MS m / z 309.0 [MH] ⁻ .

The material represented by Chemical Formula 5 is ppm (600 MHz, CD ₃ OD) 6.63 (s, H-2), 2.67 (t, J = 8.25, H-4), 2.17 (m, H-5), 5.21 ( t, J = 6.87, H-6), 1.61 (s, 7-CH ₃ ), 1.97 (t, J = 7.56, H-8), 2.08 (m, H-9), 5.14 (t, J = 6.87 , H-10), 1.60 (s, 11-CH ₃ ), 2.02 (m, H-12), 1.62 (s, H-13), 3.51 (t, J = 6.87, H-14) Resonance spectrum is shown.

The material represented by Chemical Formula 5 is ppm (600 MHz, CD ₃ OD) 176.9 (C-1), 131.1 (C-2), 145.9 (C-3), 177.4 (3-COOH), 30.5 (C-4 ), 29.2 (C-5), 126.4 (C-6), 135.7 (C-7), 16.4 (7-CH ₃ ), 41.0 (C-8), 27.8 (C-9), 125.9 (C-10 ), 135.6 (C-11), 16.2 (11-CH ₃ ), 37.1 (C-12), 32.1 (C-13), and 62.8 (C-14).

The material represented by Chemical Formula 5 was identified as novel compound SC-1 by the above result.

According to the exemplary embodiment of the present application, the schizostatin or a derivative thereof may include a material represented by the following Chemical Formula 6, but is not limited thereto;

[Formula 6]

.

.

The substance represented by Chemical Formula 6 is colorless oil, has a molecular formula of C ₂₀ H ₃₀ O ₅ , and has a molecular weight of 350.

The material represented by Chemical Formula 6 has a mass spectrometry spectrum of EIMS m / z 349.0 [M ⁻ H] ⁻ .

The material represented by Chemical Formula 6 is ppm (600 MHz, CD ₃ OD) 6.72 (s, H-2), 2.79 (t, J = 7.56, H-3), 2.18 (m, H-5), 5.16 ( t, J = 7.56, H-6), 1.60 (s, 7-CH ₃ ), 1.98 (t, J = 7.56, H-8), 2.08 (m, H-9), 5.13 (t, J = 6.87 , H-10), 1.61 (s, 11-CH ₃ ), 2.02 (t, J = 7.56, H-12), 2.13 (m, H-13), 5.39 (t, J = 6.87, H-14) , Hydrogen nuclear magnetic resonance spectra of 1.64 (s, 15-CH ₃ ) and 3.91 (s, H-16).

The substance represented by Chemical Formula 6 is ppm (600 MHz, CD ₃ OD) 169.0 (C-1), 128.4 (C-2), 148.7 (C-3), 170.2 (3-COOH), 28.9 (C-4 ), 28.8 (C-5), 124.5 (C-6), 137.4 (C-7), 16.2 (7-CH ₃ ), 40.9 (C-8), 27.8 (C-9), 125.8 (C-10 ), 136.0 (C-11), 16.3 (11-CH ₃ ), 40.6 (C-12), 27.6 (C-13), 126.8 (C-14), 135.9 (C-15), 13.9 (15-CH ₃ ) and a carbon nuclear magnetic resonance spectrum of 69.2 (C-16).

The substance represented by the said Chemical formula 6 was identified as the novel compound SC-2-1 by the said result.

According to a second aspect of the present disclosure, a step of culturing a skirt mushroom strain on a medium; Obtaining a first extract from the cultured skirt mushroom strain using a first solvent; And extracting chizostatin or a derivative thereof from the first extract using a second solvent.

Hereinafter, the manufacturing method will be described with reference to FIG. 1.

Skirt mushroom strains are cultured on a medium (S100).

According to one embodiment of the present application, the medium is PDA (potato dextrose agar), PDB (potato dextrose broth), SDA (sabouraud dextrose agar), DSA (dextrose starch agar), MM (minimal medium), CM (complete medium) , CDA (czapek dox agar) or CMC (carboxyl methyl cellulose) may be selected from, but is not limited thereto.

Preferably, after culturing the skirt mushroom strain on the PDA medium, the cultured skirt mushroom strain may be to be cultured on the PDB medium using a cork, but is not limited thereto.

Subsequently, a first extract is obtained using the first solvent from the cultured skirt mushroom strain (S200).

According to one embodiment of the present application, the first solvent may be selected from the group consisting of water, acetone, C ₁ -C ₄ alcohol or a mixed solvent thereof, and combinations thereof, but is not limited thereto.

Subsequently, chizostatin or a derivative thereof is extracted from the first extract using a second solvent (S300).

According to one embodiment of the present application, the second solvent may include an organic solvent, but is not limited thereto.

Preferably, the second solvent may be selected from the group consisting of hexane, chloroform, methanol, ethyl acetate, and combinations thereof, but is not limited thereto.

According to one embodiment of the present application, the extraction of the chyzostatin or derivatives thereof may be performed by extraction, distribution of water, or chromatography, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

Example 1 Skirt Mushroom Strain Culture and Skirt Mushroom Strain Culture Extract

Skirt mushroom strains were inoculated on PDA (potato starch 4.0 g / L, dextrose 20.0 g / L, agar 15.0 g / L) medium and incubated at 27 ° C. for 15 days. Thereafter, the mycelium at the tip of the mycelium was inoculated on PDB (potato starch 4.0 g / L, dextrose 20.0 g / L) medium using a cork borer, and cultured at 27 ° C. for 4 weeks. The skirt mushroom strain culture was soaked in acetone for 1 day. Thereafter, the skirt mushroom strain culture solution immersed in acetone was extracted at room temperature to obtain a skirt mushroom strain culture extract.

Example 2 Preparation of Chloroform Extract

Acetone was removed by concentrating the extract of Schima mushroom strain culture of Example 1. Then, the skirt mushroom strain culture extract was separated into water layer and hexane layer by partition extraction using hexane. The water layer was extracted with chloroform to separate the water layer and the chloroform layer. As a result, a chloroform extract was prepared.

Example 3 Extraction of Schizstatin or Derivatives thereof

The chloroform extract of Example 2 was subjected to silica gel chromatography using methanol as eluent. Thereafter, the eluate was subjected to MPLC using methanol at a concentration of 50% as a mobile phase to obtain a chyzostatin and a fraction. The fractions were subjected to sephadex LH-20 column chromatography using prep-HPLC and methanol at a concentration of 70% as a mobile phase to give SC-1 and SC-2-1. The fractions were subjected to silica gel chromatography using methanol as eluent. Thereafter, the eluate was subjected to sephadex LH-20 column chromatography using methanol as a mobile phase to obtain SC-3 and nerodilol.

Two-dimensional NMR spectra including ¹ H, ¹³ C, Correlation Spectroscopy (COSY) and Heteronuclear Multiple Bond Connectivity (HMBC) were measured to elucidate the chemical structure of the compounds.

Experimental Example 1

In order to investigate the antimicrobial activity against phytopathogenic fungi, the chizostatin and SC-3 of Example 3 measured the minimum inhibitory concentration (MIC), and the results are shown in Table 1.

As a test bacterium in the experiment, Rhizoctonia solani, Pyhtium sp. And the gray mold fungus Botrytis cinerea was used, the fungus was incubated for 2 to 5 days at 27 ℃.

Minimum Inhibitory Concentration [MIC (μg / ml)] Schizostatin SC-3 Rhizoctonia solani 100 100 Pyhtium sp. 100 100 Botrytis cinerea 100 ―

As a result, as shown in Table 1, Rhizoctonia solani, Pyhtium sp. It showed strong antimicrobial activity and showed antimicrobial activity against Botrytis cinerea, a gray mold.

Experimental Example 2

In order to investigate the antimicrobial activity against bacteria, the chizostatin of Example 3 was experimented using the paper disk method, and the results were shown in Table 2 by treating the sample by concentration on the disk.

In the experiment, bacteria, Staphylococcus aureus and Bacillus subtilis, were used as test bacteria, and the diameter of the growth-lowering ring (mm) was measured by culturing the bacteria at 37 ° C. for 24 hours.

Diameter of growth low ring (mm) 200 μg 100 μg 50 μg 10 μg 5 μg Bacillus subtilis 5.0 ± 0.8 3.3 ± 0.6 2.6 ± 0.1 ― ― Staphylococcus aureus 12.3 ± 0.4 8.3 ± 0.0 7.6 ± 0.1 3.2 ± 0.1 1.0 ± 0.4

As a result, as shown in Table 2 and Figure 2, chyzostatin showed a strong antimicrobial activity against Staphylococcus aureus, and showed an antimicrobial activity to Bacillus subtilis.

The above description of the present application is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present application. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present application is indicated by the following claims rather than the above description, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present application.

Claims (12)

A schizostatin or a derivative of Schizostatus strain containing the derivative thereof,
Wherein the zozostatin or derivatives thereof is an antimicrobial composition comprising a substance represented by the following formula (3):
[Formula 3]

.
delete delete delete delete delete delete Culturing the skirt mushroom strains on the medium;
Obtaining a first extract from the cultured skirt mushroom strain using a first solvent; And
Extracting chizostatin or a derivative thereof from the first extract using a second solvent
Including,
The schizstatin or a derivative thereof includes a substance represented by the following Chemical Formula 3,
The first solvent is selected from the group consisting of water, acetone, C ₁ -C ₄ alcohol or a mixed solvent thereof, and combinations thereof,
Wherein the second solvent comprises an organic solvent:
[Formula 3]

.
The method of claim 8,
The medium may be PDA (potato dextrose agar), PDB (potato dextrose broth), SDA (sabouraud dextrose agar), DSA (dextrose starch agar), MM (minimal medium), CM (complete medium), CDA (czapek dox agar) or A method for producing an antimicrobial composition, which is selected from CMC (carboxyl methyl cellulose) medium.
delete delete The method of claim 8,
Extracting the chyzostatin or derivatives thereof is carried out by extraction, distribution of water, or chromatography.

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